Endoscopic obstruction predominantly occurs in right-side colon cancer and endoscopic obstruction with tumor size ≤ 5 cm seems poor prognosis in colorectal cancer.

Background: Endoscopic obstruction (eOB) is associated with a poor prognosis in colorectal cancer (CRC). Our study aimed to investigate the association between tumor location and eOB, as well as the prognostic differences among non-endoscopic obstruction (N-eOB), eOB with tumor size ≤ 5 cm, and eOB with tumor size > 5 cm in non-elderly patients. Methods: We retrospectively reviewed the clinicopathological variables of 230 patients with CRC who underwent curative surgery. The multivariable logistic regression model was used to identify risk factors for eOB. The association between eOB with tumor size ≤ 5 cm and disease-free survival (DFS) was evaluated using multivariate cox regression analysis. Results: A total of 87 patients had eOB while 143 had N-eOB. In multivariate analysis, preoperative carcinoembryonic antigen (p = 0.014), tumor size (p = 0.010), tumor location (left-side colon; p = 0.033; rectum; p < 0.001), and pT stage (T3, p = 0.009; T4, p < 0.001) were significant factors of eOB. The DFS rate for eOB with tumor size ≤ 5 cm was significantly lower (p < 0.001) in survival analysis. The eOB with tumor size ≤ 5 cm (p = 0.012) was an unfavorable independent factor for DFS. Conclusions: The patients with eOB were significantly associated with right-side colon cancer as opposed to left-side colon cancer and rectal cancer. The eOB with tumor size ≤ 5 cm was an independent poor prognostic factor. Further studies are needed to target these high-risk groups.

Parents' Experiences of Communicating With Children About Their Diagnosis of Nonterminal Cancer and Its Related Issues: A Systematic Review of Qualitative Studies.

BACKGROUND: Many children experience serious symptoms when they are diagnosed with and treated for cancer. Through appropriate parent-child communication, parents were able to identify children's physical and psychological problems, adjust their behavior, and help them cope with the disease. OBJECTIVE: This study aimed to systematically search for and integrate evidence from qualitative studies on communication between parents and children with nonterminal cancer from parents' perspectives. METHODS: A thorough systematic review and metasynthesis of qualitative studies were conducted. Articles were searched from PubMed/MEDLINE, EMBASE, Web of Science, CINAHL, PsycINFO, and PsycArticles from the database inception to November 6, 2022. After screening and quality appraisal, 14 articles were finally included in the metasynthesis. RESULTS: Three themes and 11 subthemes were identified: (1) communication content, including diagnosis, treatment, health management, health risk, and emotion; (2) factors influencing communication, including ages of children, parents' experience of communication, parents' awareness of protection, and culture; and (3) children's responses, including acceptance and resistance. CONCLUSIONS: This systematic review found that parents were influenced by various factors during the decision-making process of parent-child communication about childhood cancer and its related issues. Parents tended to adjust their communication content and style to protect their children. IMPLICATIONS FOR PRACTICE: Future research should be conducted to explore children's experiences of communicating with their parents and analyze the similarities and differences between the communication needs of parents and children. Healthcare professionals should provide professional communication guidance to facilitate the parent-child relationship and improve the mental health of both children and their parents.

Image-activated pico-injection for single-cell analysis.

The addition of reagents into preformed droplets is a crucial yet intricate task in droplet-based applications where sequential reactions is required. Pico-injection offers high throughput and robustness in accomplishing this task, but the existing pico-injection techniques work in an indiscriminate manner, making it difficult to target particular groups of droplets. Here we report image-activated pico-injection (imgPico) for label-free, on-demand reagent supplementation into droplets. The imgPico detects the droplets of interest by real-time image analysis and makes decisions for the downstream pico-injection operation. We studied the performance of different algorithms for the image analysis and optimized the experimental settings of the imgPico. In the validation experiment, the imgPico successfully injected fluorescent dyes into droplets encapsulating one, two, and three cells, respectively, as expected. We further demonstrated the utility of imgPico by targeting droplets encapsulating single cells in droplet-based single-cell RNA sequencing (scRNA-seq) using exceedingly high cell density, and the results showed that the imgPico effectively reduced the presence of doublets in the scRNA-seq data. With the merits of being label-free and versatile, the imgPico represents a technical advance with potential applications in single-cell analysis.

Associated Clinical Factors for Coagulation Dysfunction due to Trimeresurus stejnegeri: A Retrospective Observational Study.

Background: Trimeresurus stejnegeri (T.s) accounts for most snakebites in southern China, which always leads to coagulation dysfunction. Coagulopathy due to venom is widely considered to be a characteristic phenomenon of the DIC-like syndrome. It is vitally important for first-line clinicians to improve this condition as soon as possible. However, clinical factors associated with coagulation function in Trimeresurus stejnegeri has not been well characterized yet. Materials: Patients bitten by vipers were admitted to the Emergency Department of a hospital in Shenzhen, southern China, from 2021 to 2022 and were retrospectively reviewed. Patient clinical characteristics and laboratory data were compared in the eligible patients bitten by T.s by their prothrombin time (PT), fibrinogen level (FIB), and platelet count on 2-3 days after bitten. Results: 105 patients were included in this study. The mean values of PT, FIB, and PLT are as follows: 12.8 ± 0.79 s, 2.25 ± 0.47 g/L, and 196.2 ± 57.1 × 109/L. Uric acid (UA) (367.9 ± 103.85), blood glucose (6.53 + 1.64) show negative trend of correlation, while CRP (2.12 + 4.17) shows positive trend of association with coagulation function. The smoke and systolic blood pressure may exert negative effects on PT and PLT, respectively. Logistic regression analysis indicated that uric acid (UA) shows significant connection with PT (OR = 1.15 and P value <0.0001), FIB (OR = 0.89 and P value = 0.026), and PLT (OR = 0.79 and P value = 0.007). CRP is also shown to be associated with FIB (OR = 1.33 and P value = 0.043). Conclusion: : Uric acid (UA) shows a significant association with PT, FIB, and PLT. CRP is related to FIB. Blood glucose shows a negative trend of correlation with PT. We do recommend physician should low the level of UA in some degree on the basis of injection of an antivenom serum.

Combinatorial perturbation sequencing on single cells using microwell-based droplet random pairing.

Combinatorial drug therapy reduces drug resistance and disease relapse, but informed drug combinations are lacking due to the high scale of possible combinations and the relatively simple phenotyping strategies. Here we report combinatorial perturbation sequencing (CP-seq) on single cells using microwell-base droplet random pairing. CP-seq uses oligonucleotides to barcode drugs, encapsulates drugs and cells in separate droplets, and pairs cell droplets with two drug droplets randomly on a microwell array chip to complete combinatorial drug treatment and barcode-tagging on cells. The subsequent single-cell RNA sequencing simultaneously detects the single-cell transcriptomes and drug barcodes to demultiplex the corresponding drug treatment. The microfluidic droplet operations had robust performance, with the overall utilization rate of the microwells being up to 83%. We then progressively validated the CP-seq by performing single-drug treatments and then combinatorial-drug treatments, confirming the CP-seq's capability in the collection and analysis of drug-perturbed transcriptomes. Leveraging the advantage of droplet microfluidics in massive multiplexing, the CP-seq represents a great technology for combinatorial perturbation screening with high throughput and comprehensive profiling.

Droplet Microfluidics Enables Tracing of Target Cells at the Single-Cell Transcriptome Resolution.

The rapid promotion of single-cell omics in various fields has begun to help solve many problems encountered in research, including precision medicine, prenatal diagnosis, and embryo development. Meanwhile, single-cell techniques are also constantly updated with increasing demand. For some specific target cells, the workflow from droplet screening to single-cell sequencing is a preferred option and should reduce the impact of operation steps, such as demulsification and cell recovery. We developed an all-in-droplet method integrating cell encapsulation, target sorting, droplet picoinjection, and single-cell transcriptome profiling on chips to achieve labor-saving monitoring of TCR-T cells. As a proof of concept, in this research, TCR-T cells were encapsulated, sorted, and performed single-cell transcriptome sequencing (scRNA-seq) by injecting reagents into droplets. It avoided the tedious operation of droplet breakage and re-encapsulation between droplet sorting and scRNA-seq. Moreover, convenient device operation will accelerate the progress of chip marketization. The strategy achieved an excellent recovery performance of single-cell transcriptome with a median gene number over 4000 and a cross-contamination rate of 8.2 ± 2%. Furthermore, this strategy allows us to develop a device with high integrability to monitor infused TCR-T cells, which will promote the development of adoptive T cell immunotherapy and their clinical application.

Biosynthesis of silver nanoparticle composites based on hesperidin and pectin and their synergistic antibacterial mechanism.

Silver nanoparticles (AgNPs) were widely used in the antibacterial field because of their excellent antibacterial properties. In this study, we used hesperidin and pectin as reductants and stabilizers, and prepared uniform and stable Hesperidin-Pectin AgNPs (HP-AgNPs) by a simple microwave-assisted process. Increasing the proportion of hesperidin, P-AgNPs, HP-AgNPs1, HP-AgNPs2 and H-AgNPs were obtained respectively. With the increase of hesperidin ratio, the mean particle size and zeta potential increased gradually. Fourier transform infrared spectroscopy (FTIR) analysis showed that Ag+ was reduced by hesperidin and pectin. Antibacterial tests showed that HP-AgNPs2 showed the MIC values of 66.7 µg/mL against E. coli. In addition, HP-AgNPs2 was selected to clarify its antibacterial mechanism against E. coli. Morphological experiments showed that HP-AgNPs2 stress caused damage to the cell wall of E. coli, as well as leakage of its contents and an increase in reactive oxygen species (ROS). On the other hand, the release of Ag+ during cell co-culture was studied and the results showed that most of the Ag+ released was taken up by E. coli. The synergistic effect of hesperidin and pectin resulted in a significant enhancement of the antibacterial properties of AgNPs. These preliminary data suggest that HP-AgNPs has good antibacterial activity and may be developed as an effective antibacterial nanomaterial.

Single-Cell Transcriptome Analysis of H5N1-HA-Stimulated Alpaca PBMCs.

Avian influenza A virus H5N1 is a highly pathogenic and persistently a major threat to global health. Vaccines and antibodies targeting hemagglutinin (HA) protein are the primary management strategies for the epidemic virus. Although camelids possess unique immunological features, the immune response induced by specific antigens has not yet been thoroughly investigated. Herein, we immunized an alpaca with the HA antigen of the H5N1 virus and performed single-cell transcriptome profiling for analysis of longitudinal peripheral blood mononuclear cell (PBMCs) behavior using single-cell sequencing technology (scRNA-seq). We revealed multiple cellular immunities during the immunization. The monocytes continued to expand after immunization, while the plasma cells reached their peak three days after the second antigen stimulation. Both monocytes and B cells were stimulated by the HA antigen and produced cell-type-specific cytokines to participated in the immune response. To our knowledge, this is the first study to examine the HA-specific immunological dynamics of alpaca PBMCs at the single-cell level, which is beneficial for understanding the anti-viral immune system and facilitating the development of more potent vaccines and antibodies in camelid animals.

Exome-based preconception carrier testing for consanguineous couples in China.

OBJECTIVE: To evaluate the utility of clinical exome sequencing (ES)-based carrier screening in Chinese consanguineous couples. METHODS: Consanguineous couples were screened for autosomal recessive (AR) disorders using the clinical ES of 5000 genes associated with human diseases. RESULTS: We recruited 14 couples who elected to have sequencing. One couple was related as first cousins and 13 as second cousins. Both partners carrying the same pathogenic variant were detected in four couples. One couple was found in which one partner carried a splice variant, and the other had a missence variant of the same gene. These five couples were identified as being at risk of having a child affected by an AR disorder. CONCLUSION: Our study demonstrates that ES-based preconception screening yields a clinical value for Chinese consanguineous couples. It enables to detect at-risk couples for rare AR diseases.

Survival outcomes following laparoscopic vs open surgery for non-metastatic rectal cancer: a two-center cohort study with propensity score matching.

BACKGROUND?>: Laparoscopic surgery for rectal cancer is commonly performed in China. However, compared with open surgery, the effectiveness of laparoscopic surgery, especially the long-term survival, has not been sufficiently proved. METHODS?>: Data of eligible patients with non-metastatic rectal cancer at Nanfang Hospital of Southern Medical University and Guangdong Provincial Hospital of Chinese Medicine between 2012 and 2014 were retrospectively reviewed. Long-term survival outcomes and short-term surgical safety were analysed with propensity score matching between groups. RESULTS: Of 430 cases collated from two institutes, 103 matched pairs were analysed after propensity score matching. The estimated blood loss during laparoscopic surgery was significantly less than that during open surgery (P = 0.019) and the operative time and hospital stay were shorter in the laparoscopic group (both P < 0.001). The post-operative complications rate was 9.7% in the laparoscopic group and 10.7% in the open group (P = 0.818). No significant difference was observed between the laparoscopic group and the open group in the 5-year overall survival rate (75.7% vs 80.6%, P = 0.346), 5-year relapse-free survival rate (74.8% vs 76.7%, P = 0.527), or 5-year cancer-specific survival rate (79.6% vs 87.4%, P = 0.219). An elevated carcinoembryonic antigen, <12 harvested lymph nodes, and perineural invasion were independent prognostic factors affecting overall survival and relapse-free survival. CONCLUSIONS?>: Our findings suggest that open surgery should still be the priority recommendation, but laparoscopic surgery is also an acceptable treatment for non-metastatic rectal cancer.

Reevaluation of a rightward shift in colorectal cancer: A single-center retrospective study in Tianjin.

OBJECTIVES: Some Western reports have shown a proximal shift in colorectal cancer (CRC), but there are few studies in China. This study aimed to provide more information for the management and screening of CRC by investigating trends in the anatomic distribution of CRC among the Chinese population in recent years. METHODS: A retrospective study was conducted on patients with CRC admitted to the Digestive Endoscopy Center of Tianjin Medical University General Hospital between January 2000 and December 2017. Patients were divided into a left-sided colorectal cancer (LSCRC) group and a right-sided colon cancer (RSCC) group. The detection rates of LSCRC and RSCC based on patients' age and sex, and on the time periods were analyzed. RESULTS: A total of 2319 cases were diagnosed with CRC among 75 183 consecutive patients. The prevalence of CRC showed a significant reduction from 2000-2008 to 2009-2017 (3.8% vs 2.7%, P < .001). The proportion of RSCC presented a downtrend from 2000-2008 to 2009-2017 (40.6% vs 37.7%, P > .05). There were slightly more RSCCs in female patients than in male patients, with no significant difference (40.9% vs 36.5%, P > .05). The proportion of RSCC in patients aged ≥50 years was similar to that in younger patients. The alarming symptoms between LSCRC and RSCC showed a significant difference (P < .05). CONCLUSIONS: In the present study, the prevalence of CRC declined significantly with time. However, there did not appear to be a rightward shift in CRC among the patients in Tianjin over the past 18 years.

Neuroanatomical autonomic substrates of brainstem-gut circuitry identified using transsynaptic tract-tracing with pseudorabies virus recombinants.

To investigate autonomic substrates of brainstem-gut circuitry identified using trans-synaptic tracing with pseudorabies virus (PRV)-152, a strain that expresses enhanced green fluorescent protein, and PRV-614, a strain that expresses enhanced red fluorescent protein, injecting into the rat rectum wall. 3-7 days after PRV-152 injection, spinal cord and brainstem were removed and sectioned, and processed for PRV-152 visualization using immunofluorescence labeling against PRV-152. 6 days after PRV-614 injection, brainstem was sectioned and the neurochemical phenotype of PRV-614-positive neurons was identified using double immunocytochemical labeling against PRV-614 and TPH. We observed that the largest number of PRV-152- or PRV-614-positive neurons was located in the gigantocellular reticular nucleus (Gi), lateral paragigantocellular (LPGi), rostral ventrolateral reticular nucleus (RVL), solitary tract nucleus (Sol), locus coeruleus (LC), raphe magnus nucleus (RMg), subcoeruleus nucleus (SubCD). Double-labeled PRV-614/tryptophan hydroxylase (TPH) neurons were concentrated in the RMg, LPGi and Sol. These brainstem neurons are candidates for relaying autonomic command signals to the gut. The autonomic substrate of brainstem-gut circuitry likely plays an important role in mediating different aspects of stress behaviors.

Effects of co-administration of ketamine and ethanol on the dopamine system via the cortex-striatum circuitry.

AIM: Ketamine and ethanol are increasingly being used together as recreational drugs in rave parties. Their effects on the dopamine (DA) system remain largely unknown. This study aimed to investigate the effects of consuming two different concentrations of ketamine with and without alcohol on the DA system. MATERIALS AND METHODS: We employed the conditioned place preference (CPP) paradigm to evaluate the rewarding effects of the combined administration of two different doses of ketamine (30mg/kg and 60mg/kg) with ethanol (0.3156g/kg). We evaluated the effects of the combined drug treatment on the transcriptional output of tyrosine hydroxylase (TH), dopa decarboxylase (DDC), synaptosomal-associated protein 25 (SNAP25), and vesicular monoamine transporter 2 (VMAT2) as well as protein expression level of brain-derived neurotrophic factor (BDNF) in rat prefrontal cortex (PFC) and striatum. KEY FINDINGS: We found that rats exhibited a dose-dependent, drug-paired, place preference to ketamine and ethanol associated with an elevated DA level in the striatum but not in the PFC. Moreover, treatment involving low- or high-dose ketamine with or without ethanol caused a differential regulatory response in the mRNA levels of the four DA metabolism genes and the cellular protein abundance of BDNF via the cortex-striatum circuitry. SIGNIFICANCE: This study investigated the molecular mechanisms that occur following the combined administration of ketamine and ethanol in the DA system, which could potentially lead to alterations in the mental status and behavior of ketamine/ethanol users. Our findings may aid the development of therapeutic strategies for substance abuse patients.

A new partial trisomy 12p with artery catheter vagus, congenital cataracts, external auditory canal, and no turbinate.

We describe the prenatal diagnosis and fetal phenotype of partial trisomy 12 (p12-pter) transmitted from a maternal reciprocal translocation 6;12. Genetic analysis was conducted on umbilical cord blood for a fetus accompanied with tricuspid regurgitation and orbital hypertelorism from a 27-year-old gravida 4, para 1 after sonography at gestation 35 weeks. The karyotype was unusual, with 46, XY, der (6), t (6;12) (p24; p12) mat. The pregnancy was terminated at 37 gestational weeks. The aborted fetus displayed dysmorphic features of a round flat face with prominent cheeks and high forehead, hypertelorism, short nose, broad and depressed nasal bridge, anteverted nares, deformed philtrum, open mouth, thin upper vermilion and broad everted lower lip, low-set ears and aural atresia, broad hands with simian creases, and a short neck. Fetal anatomy showed right artery catheter vagus, congenital cataract, no turbinate and external auditory canals. Through karyotype-phenotype analysis of this patient and a review of other reported cases, we believe this is a first report that expands the database of partial trisomy 12p, and is beneficial for future clinical genetic counseling. This study supports that phenotypic variability depends on the type and extent of the associated partial monosomy.